title: Cytochemical and morphometric analysis of autophagy in energy depleted rat hepatocytes.
abstract: The energy dependence of lysosomal enzyme acquisition by autophagosomes was studied in isolated rat hepatocytes by ultrastructural analysis for acid phosphatase activity. Reduction of the intracellular ATP content by addition of atractyloside or fructose decreased the flux through the autophagic proteolytic pathway to a similar extent (40-50%). Unexpectedly, in the presence of atractyloside the volume density of autophagosomes was reduced by 65%, whereas in the presence of fructose this reduction was only 20%. The volume density of lysosomes was not significantly affected by either of the two compounds. It is concluded that partial ATP depletion by fructose not only inhibits sequestration of cytoplasmic material in autophagosomes, but also affects the fusion between autophagosomes and lysosomes. Since fructose, in contrast to atractyloside, does not affect the cytosolic phosphate potential, it is proposed that autophagic sequestration is more sensitive to changes in the cytosolic phosphate potential whereas the fusion between autophagosomes and lysosomes is more responsive to changes in the ATP concentration.
authors.affiliation: Laboratory of Cell Biology and Histology, University of Amsterdam, The Netherlands.
authors.forename: J P
authors.initials: JP
authors.lastname: Schellens
authors.forename: H
authors.initials: H
authors.lastname: Vreeling-Sindelárová
authors.forename: P J
authors.initials: PJ
authors.lastname: Plomp
authors.forename: A J
authors.initials: AJ
authors.lastname: Meijer
mesh_terms: D000135:Acid Phosphatase / Q000032:analysis; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001278:Atractyloside / Q000494:pharmacology; D001343:Autophagy; D002478:Cells, Cultured; D004734:Energy Metabolism / Q000187:drug effects; D005508:Food Deprivation; D005632:Fructose / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D001278:Atractyloside; D005632:Fructose; D000255:Adenosine Triphosphate; D000135:Acid Phosphatase